Plum Island Ecosystems LTER Database

Acceptance and utilization of LTER data requires that:

(1) The Principal Investigator be sent a notice stating reasons for acquiring any data and a description of the publication intentions.
(2) The Principal Investigator of the data set be sent a copy of the report or manuscript prior to submission and be adequately cited in any resultant publications.
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Benthic chlorophyl measurements of surface sediments (top 0-2cm) from a variety of habitats in saltmarsh tidal creeks of the Rowley River, Rowley and Ipswich, MA. Habitats include: mudflat, filamentous algae, tall Spartina alterniflora, short Spartina alterniflora and Spartina patens zones. The TIDE project aims to simulate eutrophication on a large scale by the addition of NO3- aiming to reach 70µM concentrations from May to September every year during the growing season. This fertilization of the marsh has been going on at Sweeney Creek since the 2004 growing season through 2011 and at Clubhead Creek in 2005 and then from 2009 till 2011.

Version 01: January 27, 2012, updated data and metadata. Used MarcrosExportEML_HTML (working)pie_excel2007.xlsm 1/18/12 4:51 PM for QA/QC to EML 2.1.0

Version 02: June 20, 2013 data and metadata updated to comply with importation to Drupal and LTER PASTA. Research location name and description updated.Used MarcrosExportEML_HTML (working)pie_excel2007.xlsm 3/14/13 12:02 PM for QA/QC to EML 2.1.0

For each transect collect the following number of samples in each habitat: 3 mudflat, 3 filamentous algae, 3 tall Spartina alterniflora, 3 Spartina patens, 3 short Spartina alterniflora

Habitat:Collect samples at low tide in order to access MF, FA, and TSA habitat types.Use transect as guideline for picking sample locations, but do not work directly along the transect line.Choose general sampling location and toss syringe to “haphazardly” pick exact spot.To collect sample: slowly push down syringe while pulling up on the plunger (try not to compress the sample). Try to get as far down as possible and then slowly pull the tube out of the substrate. Pull the plunger out and stick it into the opposite end of the syringe. Using ruler to measure, push out 2 cm of sample into the appropriately labeled centrifuge tube. (Be careful to measure two centimeters as carefully as possible each time. Calculations of chlorophyll after analysis assume equal volumes of sediment in each sample.)MF: Take sample in exposed mud flats along creek channel during low tide. Avoid inserting syringe into an area with root masses and fibers, it is generally very easy to sample in the mud flats.FA: Take sample along creek wall below TSA roots layer.TSA: Same as mudflats.SP & SSA: Sampling is more difficult, because of the dense roots. Cut around cylinder with knife as you are inserting syringe into ground. Once syringe is inserted far enough cut a cone shape underneath syringe with knife and pull out syringe at an angle in order not to lose the sample.Collection takes about a half day per creek with two people.Once samples are collected, wipe off as much mud on the outside of the tubes and store in labeled plastics bags. Samples must remain frozen until analysis.

*adapted from S. Kelsey 2001 protocolupdated 12/20/07 CGK

LAB EXTRACTIONStep 1: Extraction (typically done in the pm the day before analysis on spec)Supplies:50-ml polypropylene centrifuge tubes containing sample (frozen and in the dark)100% acetone90% acetoneCoolers with iceRe-pippeterLatex (non-nitrile) gloves

Considerations:Chlorophyll degrades easily. As much as possible, keep samples cool and dark.Work in a hood to minimize your exposure to acetone fumes and cap samples tightlywhen not in use. Acetone eats many plastics so be mindful of the materials you are usingto store and transfer samples. Acetone will also wash away sharpie- so keep track ofsample numbers and protect your labels. Avoid using acid-washed equipment tominimize potential acid-contamination of samples. Length of extraction should bearound 16 hours, so plan to time your sample prep and analysis accordingly. Prep onlythe amount of samples you can feasibly analyze in one sitting. Usually no more than 40-50 samples at a time.

Prep samples with acetone:

1. Allow samples to thaw at room temperature for about two hours before putting incoolers. Samples should be thawed, but cool to the touch.2. Clean any excess mud off of sample containers and transfer thawed samples tocoolers with ice3. Store acetone solution in coolers on ice4. Under hood, use a re-pippeter to add 25mls of 100% acetone to each tube5. Shake each sample briefly by hand to break up sediment at the bottom of eachtube6. Place tubes back on ice in coolerSonication:

1. Sonicator is in general-use equipment room of Lillie building (2nd floor). ContactHerb Luther (x7722) or Louie Kerr (x7273) for questions regarding the setup ofthe sonicator. Sonicator may be in TIDE lab in the 1st floor of ESL.2. Bring cooler with samples, kimwipes, gloves, stopwatch, and plastic beaker3. Wear ear protection when using the sonicator and place a sign on the door tonotify others of its use.4. Place sample tube in plastic beaker with ice, un-cap and sonicate at setting 7(microtip max) for 30 seconds, looking for signs of boilage. Avoid touching theprobe to the sides of the sample tube or removing the probe from the liquid whilerunning.

Reference creeks are West and NelsonNutrient Enriched creeks are Sweeney and Clubhead. Sweeney has been fertilized every summer from 2004-2011. Clubhead had been fertilized in 2005 and then from 2009-2011.

All creeks have two branches determined by direction at confluence while facing upstream (Left or Right).

Plant transects run perpendicular to creeks at various points with a front pole at the creek bank and a back pole an the high marsh. Transects are in the order of 1, 2, 4, 3 or blue, green, red, yellow from confluence of branch.